The solubilization of myofibrillar proteins of vertebrate skeletal muscle in water

Myofibrillar proteins of vertebrate skeletal muscles are insoluble in solutions of ionic strength that approximate physiological conditions. We established a method to solubilize more than 80% of chicken breast muscle myofibrillar proteins in water for the use of meat as a source of food protein. SDS‐polyacrylamide gel electrophoretic patterns of water‐soluble myofibrillar proteins demonstrated that all identified myofibrillar proteins except connectin/titin were soluble in water. A part of α‐actinin was released from myofibrils by repeated washing with 2.5 mmol/L NaCl and 5 mmol/L L‐histidine solution, and subsequent destruction of connectin/titin in washed myofibrils by ultrasonication resulted in solubilization of a large fraction of chicken breast muscle myofibrillar proteins in water. Myofibrillar proteins of chicken leg, pork loin, beef shoulder loin, and lamb were also solubilized in water using this procedure.     

Proteome analysis of whole and water‐soluble proteins in masseter and semitendinosus muscles of Holstein cows

To assess both quantitative and qualitative differences between the slow‐ and fast‐type muscles, masseter (slow) and semitendinosus (fast) from four Holstein cows were analyzed by two‐dimensional difference gel electrophoresis (2D DIGE) and mass spectrometry. The proteome analysis identified 27 spots as 20 proteins in the whole protein fraction extracted with 8 mol/L urea solution, and 16 spots were identified as 11 proteins in the water‐soluble protein fraction. Two slow‐type myofibrillar proteins (myosin light chain‐1 slow‐b and myosin light chain‐2 slow), and aconitase‐2 mitochondria were present at higher levels in the masseter muscle (P < 0.05). Four fast‐type myofibrillar proteins (myosin light chain‐1 fast, myosin light chain‐2 fast, myosin light chain‐3 fast and tropomyosin‐1), and three enzymes of glycolytic pathway (enolase‐3, aldolase‐A and triosephosphate isomerase), were present at higher levels in the semitendinosus muscle (P < 0.05). Our proteome analysis showed that the composition of sarcoplasmic proteins as well as myofibrillar proteins was clearly different between slow‐ and fast‐type muscles.     

Improvement of the gelling properties of meat emulsion gel by the addition of porcine sarcoplasmic proteins

The effects of porcine sarcoplasmic proteins (SP) on the physicochemical properties of meat emulsion gel were examined. Meat emulsion was prepared from water‐washed pork meat (WWM), corn oil and SP. Whole SP (W‐SP) enhanced the breaking stress of the WWM emulsion gel as well as other animal proteins in the presence of 0.2 mol/L NaCl. The breaking stress of the emulsion with W‐SP increased with an increase in corn oil content. Furthermore, this tendency was more noticeable at a lower NaCl concentration (0.15 mol/L) rather than at 0.45 mol/L NaCl. Ammonium sulfate (AS) treatment fractionated W‐SP into three portions (SP‐f1, SP‐f2 and SP‐f3), which were the precipitates at 0–50% and 50–75% AS saturation and the supernatant at 75% AS saturation, respectively. The fractions SP‐f2 and SP‐f3 increased the gel strength more than W‐SP. In particular, the fraction SP‐f3 increased the gel strength approximately 10‐fold compared to the control. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis analysis showed that SP‐f3 had several kinds of proteins and a main protein with a molecular mass of 35 kDa, which corresponded to glyceraldehyde 3‐phosphate dehydrogenase. These results indicate that the influence of SP should not be ignored when processing of low‐salt meat products and the fraction SP‐f3 has a gel‐enhancing factor for myofibrillar proteins.     

Physicochemical properties of the thermal gel of water-washed meat in the presence of the more soluble fraction of porcine sarcoplasmic protein

We investigated the physicochemical properties of the thermal gel of water‐washed pork meat (WWM) in the presence of the soluble fraction of porcine sarcoplasmic protein (SP) obtained with ammonium sulfate at 75 percent saturation. Two precipitated fractions of SP were obtained at 0–50 percent and 50–75 percent saturation, named SP‐f1 and SP‐f2, respectively, and the soluble fraction obtained at 75 percent saturation, SP‐f3, was used. Sodium dodecyl sulfate‐polyacrylamide gel electrophoresis showed that SP‐f3 contained mainly glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH), while SP‐f1 and SP‐f2 had other SPs such as phosphorylase b, enolase, actin and phosphoglycerate mutase. The gel strength of WWM was greater when SP‐f3 rather than one of various animal proteins such as bovine plasma (BP), egg white, or whey protein isolates (WPI), was added and SP‐f3 had a gel‐enhancing effect as good as that of polyphosphate (PP). The gel strength of WWM with added SP‐f3 increased significantly with NaCl at 0.15 mol/L or more, but not in the absence of NaCl (0 mol/L). The effect of SP‐f3 was evident at neutral pH and maximum gel strength was obtained at a pH above 6.0. Differential scanning calorimetric (DSC) analysis showed that an endothermic peak corresponding to myosin heads in WWM shifted to a lower temperature with the addition of SP‐f3, as in the case of PP, though there was no such shift in the presence of other animal proteins (BP, egg white and WPI), suggesting that SP‐f3 increases the gel strength of WWM through the dissociation of actomyosin similar to PP. Scanning electron microscopy (SEM) revealed wall‐like structures among the protein strands in the WWM gel matrix in the presence of SP‐f3. The results of DSC and SEM indicated that the formation of a gel network in meat products is reinforced with GAPDH in SP after the interaction between GAPDH and myofibrillar protein.     

Myofibrillar proteolysis in chick muscle cell cultures during heat stress

The effect of heat stress on protein oxidation and myofibrillar proteolysis in chick myotubes was investigated. Myotubes were incubated at 37 or 41°C for 6 and 24 h. Protein carbonyl content, as an index of protein oxidation, increased more at 41°C than at 37°C for 6 and 24 h incubations. N^τ‐methylhistidine release as an index of myofibrillar proteolysis also increased more at 41°C than at 37°C for 6 and 24 h incubations. Proteasome activity also increased more under those same conditions. Calpain and cathepsin D but not B + L activities showed a greater increase at 41°C than at 37°C for 24 but not the 6 h incubation. These results indicate that heat stress increases protein oxidation and proteasome activity, resulting in an increase in myofibrillar proteolysis for short‐term incubation and, for long‐term incubation, it increases calpain, proteasome and cathepsin D activities, finally accelerating myofibrillar proteolysis in chick myotubes.     

Effect of porcine stress syndrome on the solubility and degradation of myofibrillar/cytoskeletal proteins.

This study examined the effect of stress classification (stress-positive, stress-carrier, stress-negative) of pigs on selected properties of postmortem muscle, including protein solubility and degradation of proteins such as titin. Longissimus muscle samples were removed 45 min postslaughter, divided into samples, and stored at 0 to 2 degrees C for analysis at 0, 1, 3, 5, and 7 d postmortem. Whole-muscle samples (homogenates) and purified myofibrils were prepared from each sample for analysis by SDS-PAGE. A portion of each muscle sample also was extracted 1) with a low-ionic-strength solution to obtain a sarcoplasmic protein fraction and 2) with two different high-ionic-strength solutions to obtain a myofibrillar/cytoskeletal protein fraction for measurement of protein solubility and for analysis of extracts by SDS-PAGE. No significant differences were observed between muscle from stress-negative and stress-carrier animals in this study. Sarcoplasmic (P less than .05) and myofibrillar/cytoskeletal (P less than .01) protein solubility was lower in muscle samples from stress-positive animals than in muscle samples from stress-carrier and stress-negative animals at all postmortem times studied. The high molecular weight protein titin was degraded more slowly postmortem in muscle from stress-positive than in muscle from stress-negative animals, as observed by SDS-PAGE analysis of whole-muscle samples (homogenates) an myofibrils. The combination of lowered protein solubility and reduced rate of postmortem degradation of structural proteins such as titin may explain, at least in part, the reduced quality and protein functionality of muscle from stress-positive pigs.     

土壤铜形态与环境因素的关系及其对黑麦草生长的影响

采用盆栽试验研究酸度与氮素水平对土壤易溶态铜形态的影响,以及土壤铜含量与形态对黑麦草生长的影响。结果表明,土壤水溶态铜、有机碳结合态铜、离子态铜与铜处理水平呈极显著正相关关系,相关系数分别达到0.70、0.96、0.61。土壤酸度影响土壤中各形态铜的含量,随pH值降低土壤水溶态铜、离子态铜均显著增加,在pH4.7土壤上二者的含量随铜处理浓度的不同分别为pH6.0土壤上的1.6～6.3倍和39～148倍,两种土壤上水溶态铜、离子态铜含量差异达显著水平。铜处理水平、水溶态铜、有机碳结合态铜、离子态铜含量均强烈影响黑麦草的生长和其对铜的吸收,牧草鲜重与各形态铜呈极显著负相关,相关系数分别为-0.73、-0.76、-0.71、-0.72,而植株铜含量则与各形态铜间呈极显著正相关关系,相关系数分别为0.61、0.87、0.56、0.87。土壤氮素水平影响土壤溶解有机碳的含量,但对土壤各种铜的形态及牧草对铜的吸收并无显著影响。     

Effect of fast freezing then thaw‐aging on meat quality attributes of lamb M. longissimus lumborum

The objective of this study was to determine the effect of two different freezing rate then thaw‐aging regimens on the quality attributes of lamb loins. The loins were randomly allocated to one of five different freezing/thawing/aging regimes: fast‐(FF1A0) and slow‐(SF1A0) frozen only; fast‐(FF1A2) and slow‐(SF1A2) frozen then thaw‐aged for 14 days; aged for 14 days never frozen (A2). FF1A2 samples had a significantly higher water‐holding capacity compared to the slow frozen regardless of further aging periods. FF1A2 samples had lower (p < 0.05) shear force values than A2 and higher (p < 0.05) water‐holding capacity compared to the SF1A2. Fast freezing resulted in more intracellular cryo‐damage, whereas slow freezing resulted in extracellular cryo‐damage. FF1A0 and SF1A0 samples had lower (p < 0.05) myofibrillar proteins degradation. This study demonstrated that fast freezing then thaw‐aging can result in an improved water‐holding capacity and tenderness through the minimization of extracellular ice crystal formation, reduction in purge and drip losses, and improved proteolysis in thawed lamb.     

Physico‐chemical characteristics of Longissimus lumborum muscle in goats subjected to halal slaughter and anesthesia (halothane) pre‐slaughter

This study assessed the effect of halal slaughter and anesthesia pre‐slaughter followed by bleeding on meat quality characteristics of goats. Eleven male Boer cross goats were divided into two groups and subjected to either halal slaughter (HS) or anesthesia with halothane and propofol pre‐slaughter (AS). At pre‐rigor, HS had significantly lower (P < 0.05) muscle pH and glycogen than AS. However, no significant difference was observed in the pH and glycogen content between the treatments on 1, 3 and 7 days post mortem. The drip loss of HS was significantly lower (P < 0.05) than that of AS at all aging periods. Treatment had no effect on sarcomere length, myofibrillar fragmentation index and shear force values, loss of thiol groups and degradation of major myofibrillar proteins. It can be concluded that HS did not have deleterious effect on meat quality traits of goat when compared to AS.     

Effect of calpastatin on degradation of myofibrillar proteins by mu-calpain under postmortem conditions.

To improve our understanding of the regulation of mu-calpain activity in situ during postmortem storage of muscle, the effect of different calpastatin levels on proteolysis of myofibrillar proteins by mu-calpain in a system closely mimicking postmortem conditions was studied. Increasing the amount of calpastatin in the incubations limited both the rate and extent of proteolysis of myofibrillar proteins and autolysis of mu-calpain. Excess calpastatin (i.e., a mu-calpain:calpastatin ratio of 1:4) did not inhibit proteolysis completely. Western blot analysis revealed that proteolysis of myofibrillar proteins virtually ceased after 7 d of incubation, despite the presence of partly autolyzed, therefore seemingly active, mu-calpain. A series of incubations of autolyzed mu-calpain revealed that the autolyzed form of this enzyme is unstable at an ionic strength observed in postmortem muscle. The possible significance of these results in terms of the regulation of mu-calpain activity in postmortem muscle is discussed.     

Effect of pH and ionic strength on mu- and m-calpain inhibition by calpastatin

The objectives of this study were to determine the extent to which pH and ionic strength influence mu- and m-calpain activity and the inhibition of calpains by calpastatin. Calpastatin, mu-calpain, and m-calpain were purified from at-death porcine semimembranosus. Mu-calpain or m-calpain (0.45 U) were incubated with the calpain substrate Suc-Leu-Leu-Val-Tyr-7-amino-4-methyl coumarin in the presence of calpastatin (0, 0.15, or 0.30 U of calpain inhibitory activity) under the following pH and ionic strength conditions: pH 7.5 and 165 mM NaCl or 295 mM NaCl; pH 6.5 and 165 mM NaCl or 295 mM NaCl; and pH 6.0 and 165 mM NaCl or 295 mM NaCl. The reactions were initiated with addition of 100 microM (mu-calpain) or 1 mM CaCl2 (m-calpain), and calpain activity was recorded at 30 and 60 min. Mu-calpain had the greatest (P < 0.01) activity at pH 6.5 at each ionic strength. Higher ionic strength decreased mu-calpain activity (P < 0.01) at all pH conditions. Inhibition percent of mu-calpain by calpastatin was not affected by pH; however, it was influenced by ionic strength. Inhibition of mu-calpain by calpastatin was higher (P < 0.01) at 295 mM NaCl than at 165 mM NaCl when 0.3 units of calpastatin were included in the assay. Activity of m-calpain was greater (P < 0.01) at pH 7.5 than at pH 6.5. m-Calpain activity was not detected at pH 6.0. Inhibition of m-calpain was greater (P < 0.01) when 0.15 and 0.3 U calpastatin were added at pH 6.5 than 7.5 at 165 mM NaCl, whereas percentage inhibition of m-calpain was greater (P < 0.01) at 295 mM than 165 mM NaCl at pH 7.5 and 6.5. These observations provide new evidence that defines further the influence of pH decline and increased ionic strength on mu-calpain, m-calpain, and calpastatin activity, thereby helping to more accurately define a role for these enzymes in the process of postmortem tenderization.     

Use of chemical characteristics to predict the relative bioavailability of supplemental organic manganese sources for broilers

Twelve organic Mn sources and MnSO4 were evaluated by polarographic analysis and via solubility in buffers (pH 5 and 2) and deionized water. Fractions from solubility tests were evaluated by gel filtration chromatography for structural integrity. Organic Mn sources included five Mn methionine complexes (Mn Met A to Mn Met E), two Mn proteinates (Mn Pro A and Mn Pro B), and five Mn amino acids (Mn AA A to Mn AA E). Sources varied considerably in chemical characteristics. Chelation strength (Qf) ranged from weak (1.9 Qf-values) to strong complexes (115.4 Qf-values). No complexed Mn was found in filtrates at pH 2.0 or 5.0. A 42-d bioassay was used to estimate relative bioavailability of Mn sources for chicks fed diets supplemented with 60, 120, or 180 mg Mn/kg. Bone Mn, heart Mn, heart manganese-superoxide dismutase activity (MnSOD), and heart MnSOD mRNA increased (P < 0.001) as dietary Mn increased. Only heart MnSOD mRNA tended (P < 0.10) to differ among dietary Mn sources. For bioassays of Mn, the MnSOD mRNA level in heart was more sensitive than the MnSOD activity in heart or other indices. Relative to MnSO4 (assigned 100%), slope ratios of MnSOD mRNA levels in heart gave bioavailabilities of 99, 132, and 113% for Mn Met E, Mn AA B, and Mn AA C sources with weak, moderate, and strong chelation strength, respectively. The bioavailability of Mn was more closely related to chelation strength as measured by polarography than to chemical traits assessed by solubility or structural integrity.     

Influence of transport conditions and pre‐slaughter water shower spray during summer on protein characteristics and water distribution of broiler breast meat

This study investigated the effects of pre‐slaughter transport during summer and subsequent water shower spray on broiler meat quality and protein characteristics. Arbor Acres broiler chickens (n = 126, 42 days old, mixed sex, 2.5‐3 kg) were randomly categorized into three treatments: (i) control group without transport (C); (ii) 30 min transport (T); and (iii) 30 min transport followed by 10 min water shower spray and 20 min lairage (T/W). Each treatment consisted of six replicates with seven birds each. Ambient temperature was 32‐35°C during transportation. Results indicated that transport during high ambient temperature denatured myosin and sarcoplasmic proteins, led to decreased protein solubility and resulted in glycogen phosphorylase precipitated to the myofibrillar fraction. Furthermore, meat quality in the transport group showed a pale, soft and exudative (PSE)‐like syndrome. Water shower spray during lairage after transport reduced the degree of protein denaturation and lessened the deterioration of meat quality.     

Post mortem development of meat quality as related to changes in cytoskeletal proteins of chicken muscles

1. A procedure was developed to separate high and medium molecular weight myofibrillar proteins from chicken muscular tissue with a high resolution by flat bed sodium-dodecyl-sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and subsequent detection by either a general protein stain or Western blotting. These procedures were used to analyse the degradation process of cytoskeletal proteins in chicken breast and leg muscles during meat ageing. 2. This study demonstrates the degradation of all the examined cytoskeletal proteins: titin, nebulin and desmin as well as vinculin, a protein component of the costamere structure. All the examined proteins were found to be degraded during ageing of chicken breast and leg muscles. 3. Degradation of titin, nebulin and desmin started at 3 h post mortem in breast muscle. Intact titin and nebulin disappeared within 1 d. Intact desmin and vinculin were not detectable after 3 d post mortem. In leg muscle, the degradation process of all the examined proteins evolved much more slowly than in breast chicken muscles. 4. The changes observed in shear force, myofibrillar fragmentation and cooking loss were related to changes in cytoskeletal proteins and used to identify marker proteins or degradation products for the purpose of monitoring the development of meat ageing. The ageing process was faster in breast muscle than in leg muscle. 5. Significant correlations were found between degradation processes of titin, nebulin, and desmin and shear force, as well as myofibril fragmentation index of breast and leg muscles.     

The scientific basis for establishing solubility criteria for veterinary species

The relationship between factors influencing the in vitro and in vivo drug solubilization is highly dependent upon the physiology of the individual animal species and the physico-chemical properties of the drug. The solubilization of a drug in an aqueous medium is controlled by the strength of the interaction between a molecule of the active pharmaceutical ingredient (API) and the molecules of the solvent. In this regard, the stronger this interaction, the greater the likelihood that the drug will go into solution. Counteracting this solubilization process is the strength of the affinity of the solute for itself, or how tightly bound the compound is to its own solid state form. The stronger affinity of a solute for itself, the more difficult it will be for that molecule to go into solubility. However, solubility is not a universal value. Rather, it should be considered from the perspective of the interactions between the API in its solid form and the solution conditions such as: pH, the nature of the primary solvent, the presence of co-solvents, the presence of solubilizing additives, the ionic strength of the medium, incubation time, and temperature. Each of these factors needs to be considered when evaluating potential interspecies differences in drug solubility.     

Feeding hay rich in water‐soluble carbohydrates improves ruminal pH without affecting rumination and systemic health in early lactation dairy cows

Forages rich in water‐soluble carbohydrates (WSC) might be an ideal energy source during early lactation, as they provide both energy for milk production and structural fibre to promote chewing and rumen buffering. Thus, the aim was to investigate feeding strategies based on high‐quality hay rich in WSC with graded amounts of concentrate on ruminating behaviour, ruminal pH and systemic health variables. Twenty‐four Simmental cows were randomly allocated to four groups beginning 10 days before until 28 days after calving. Diets were 60LQH (60% fibre‐rich hay plus 40% concentrate), 60HQH, 75HQH and 100HQH (60%, 75% and 100% high‐quality hay, plus 40%, 25% and 0% concentrate, respectively). Hay qualities differed in contents of WSC (110 g vs. 198 g of dry matter [DM]), neutral detergent fibre (646 g vs. 423 g of DM) and crude protein (65 g vs. 223 g of DM). Rumination was recorded using the Rumiwatch system over 4 days during the last week. Weekly serum samples were analysed for the liver enzymes aspartate aminotransferase, glutamate dehydrogenase and γ‐glutamyltransferase and the acute phase proteins serum amyloid A and haptoglobin. Four cows per group received a wireless pH sensor orally placed into the rumen one week before the expected calving date. Daily time spent chewing did not differ between groups. Likewise differences in minimum, maximum and mean pH‐values were not significant, but daily time of reticular pH <6 was longer in cows fed 60LQH compared to cows fed 100HQH (p = 0.043) and in tendency to cows fed 75HQH or 60HQH (p = 0.072 or p = 0.086, respectively). Blood parameters were unaffected by diet. Accordingly the present results demonstrate that feeding hay rich in WSC helped stabilizing the reticuloruminal pH in early lactation dairy cows, even in combination with 40% concentrates in DM.     

An ELISA for detection of antibodies against porcine epidemic diarrhoea virus (PEDV) based on the specific solubility of the viral surface glycoprotein.

Viral proteins of porcine epidemic diarrhoea virus (PEDV) were extracted from the cytoplasm of infected Vero cells using hypotonic conditions and a non-ionic detergent. Both the pH and the NaCl concentration of the extraction buffer were varied in attempts to increase the solubility of the virion spike glycoproteins (S-protein) and of the nucleocapsid proteins (N-protein). Monoclonal antibodies, hyperimmune sera and convalescent pig sera were used to identify and monitor these proteins by immunoprecipitation and Western blots. The solubility of the S-protein was optimal at pH 4, whereas that of the N-protein was optimal at pH 9. Consequently, it was possible to enrich for either S-protein or N-protein; increases in the NaCl concentration of the buffer were of no advantage in this respect. Enriched preparations of the S-protein and N-protein were used as ELISA antigen for the S-ELISA and N-ELISA, respectively. The S-ELISA proved to be the more effective of the two immunoassays. Antibodies against S-protein remained detectable for longer periods of time than anti-N-protein antibodies in the sera of PEDV-infected pigs. Using this ELISA of increased sensitivity, it was observed that only a small number of farms in Switzerland had been infected with PEDV.     

Comparison of proteolysis in native, heat-treated and aged proteins from turkey meat

1. The present study compared the ability of native, heat-treated and aged turkey breast muscle proteins to undergo proteolysis by digestive tract proteases. 2. Domestic turkey toms were slaughtered under laboratory conditions. Breast muscles were excised immediately post mortem; one was placed under conditions to develop exudative meat by maintaining the muscle at 40 degrees C for at least 30 min and the other was refrigerated under commercial conditions. 3. Meat was collected and stored for 7 d at 4 degrees C. Breast samples removed at d 0 and d 7 were frozen and stored at -80 degrees C until used for determination of solubility, protein surface hydrophobicity and protein oxidation through carbonyl content. Measurements of pepsin and trypsin/chymotrypsin activities were performed in vitro on myofibrillar proteins. 4. Storage increased carbonyl content in control samples while the oxidation increase was not significant in heat-treated myofibrillar protein. Hydrophobicity was not affected by storage time or treatment or protein solubility. 5. Storage significantly increased trypsin + chymotrypsin activity only in the control group. The activities of pepsin and trypsin + chymotrypsin were negatively correlated with protein surface hydrophobicity.     

The influence of crossbreeding on collagen solubility and tenderness of Infraspinatus and Semimembranosus muscles of semi‐intensively reared young bulls

The aim of the present study was to investigate the effects of crossbreeding on collagen content and solubility, shear force (WBSF) and the eating quality of Infraspinatus (INF) and Semimembranosus (SM) muscles of young bulls and the relationships between collagen content and solubility, shear force and the eating quality of beef. The experimental material comprised muscles of crossbred young bulls (about 600 days old) of Polish Holstein‐Friesian (PHF) × Limousine (LM) (n = 10), PHF × Charolaise (CH) (n = 9), PHF × Hereford (HER) (n = 9) breeds. The crossbreeding influenced WBSF, aroma and taste, total, water‐soluble, acid‐soluble, total soluble and insoluble collagen content, as well as the acid‐soluble, total soluble and insoluble collagen proportions. WBSF was significantly negatively correlated with sensorial tenderness and water‐soluble collagen content. The eating quality of beef obtained from different crossbreds was similar; however, the meat from PHF × LM and PHF × HER bulls had lower WBSF values than PHF × CH bulls. © 2016 Japanese Society of Animal Science     

Effect of stresses before slaughter on changes to the physiological, biochemical and physical characteristics of duck muscle.

1. This experiment was designed to study the effects of fasting and enforced exercise on the physiological, biochemical and physical characteristics of duck muscle. 2. Sixty 75-d-old male ducks weighing 3.0 +/- 0.2 kg were assigned to three treatments: a control, and an 8 and 24 h fast plus enforced exercise for 10 min. The ducks were then sacrificed and the carcass stored at 4 degrees C for 24 h. 3. Although the pH and serum lactate contents gradually increased with fasting time the responses were not significant. The ultimate pH was elevated and the lactate of breast and thigh muscles was lower in stressed birds. 4. The activity of lactic dehydrogenase was significantly increased by the stress, and the activities of creatine phosphokinase and alkaline phosphatase were also increased slightly. However, no effect was found on the ATPase activity of the myofibrillar protein of either breast or thigh muscle as a result of the stress. The ATPase activity of myofibrillar protein of breast muscle significantly increased with storage time. 5. The extractability of myofibrillar protein increased with storage time for all treatments. The SDS-PAGE patterns of myofibrillar proteins were also studied. 6. Consequently DFD-like muscle was observed in the breast and thigh muscles of ducks which had been stressed.